丙酮酸乙酯对热打击后血管内皮细胞增殖活性的影响

Effects of ethyl pyruvate on proliferation activity of vascular endothelial cells after heat stress

目的探讨丙酮酸乙酯(EP)对热打击后血管内皮细胞增殖活性的影响。方法将人脐静脉血管内皮细胞(HUVECs)分别置入不同温度设置(39℃,41℃,43℃)的细胞培养箱内进行热打击4 h,或放入相同温度设置(43℃)的细胞培养箱内接受热打击不同时间(2 h,3 h,4 h)(时间点选取的依据——以上时间点是根据预实验的结果来选择的,预实验中43℃持续热打击5 h时细胞大部分都出现坏死脱壁,崩解成细胞碎片),对照组(CONT)则始终置于37℃细胞培养箱中培养,然后在倒置显微镜下观察各组细胞的形态学变化,并采用CCK-8试剂盒检测各组细胞的增殖活性;根据上述实验结果选取合适的热打击温度(43℃)及时间点(4 h),予以浓度为10 mmol/L的EP进行干预,然后采用CCK-8试剂盒检测细胞的增殖活性。结果随着热打击温度不断升高或热打击时间不断延长,镜下细胞的形态逐渐发生变化,以43℃热打击4 h组(HS组)细胞的形态变化最为明显;另外随着热打击温度的不断升高和热打击时间的不断延长,细胞的增殖活性逐渐降低,且与对照组(CONT)相比,均以43℃热打击4 h组细胞的增殖活性降低最为显著(F=25.79,P<0.001),而且与热打击组(HS)相比,EP干预组(HS+EP)细胞的增殖活性明显增高(P<0.001)。结论EP可以明显减轻热打击对HUVECs增殖活性的影响,有助于缓解高热所造成的血管内皮细胞活性的改变。

Objective To investigate the effect of ethyl pyruvate on proliferation activity of vascular endothelial cells after heat stress.Methods Human umbilical vein endothelial cells(HUVECs)were placed in incubators with different temperatures(39℃,41℃,43℃)for heat shock for 4 h,or placed into the incubator with the same temperature at 43℃and received heat shock for different times(2 h,3 h,4 h),and the control group were always placed in 37℃incubator.Then the morphological changes of the cells were observed under an inverted microscope,and the cell proliferation activity was detected by the cell counting-8(CCK-8)kit.According to the above experimental results,the optimal intervention temperature(43℃)and time point(4 h)of heat stress were selected.After that,ethyl pyruvate(EP)with a concentration of 10μmol/L was used for intervention(HS+EP group),and the cell proliferation activity was detected by CCK-8.Results With the increasing of the heat stress temperature or the extension of exposure time,the cell morphology gradually changed under the inverted microscope,and the cells in 43℃incubator for 4 h was the most obvious;and the cell proliferation activity of HUVECs decreased gradually,and the most significant decrease occurred in the group that exposure for 4 h in 43℃(F=25.79,P<0.001 vs.control group).In addition,the cell proliferation activity of HUVECs in the HS+EP group was significantly higher than that in the HS group(P<0.001).Conclusions EP can reduce significantly the effect of heat stress on the proliferation activity of HUVECs,and help to alleviate the changes of vascular endothelial cell activity caused by heat stress.